Method for detecting cell proliferative disorders

a cell proliferative disorder and detection method technology, applied in the field of detection of target nucleic acid sequences, can solve the problems of difficult detection and treatment of bladder cancer, expensive and invasive procedures, and inability to detect and treat bladder cancer, etc., to achieve sensitive and non-invasive screening, fast and reliable results

Inactive Publication Date: 2009-11-19
THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE
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  • Abstract
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Benefits of technology

[0019]The invention also provides a fast, reliable, sensitive and non-invasive screening method for detecting genetic instability of microsatellite DNA. An amplification or deletion of the small tandem repeat DNA sequences indicates genetic instability in the microsatellite DNA that is at or corresponds to an allele.

Problems solved by technology

Cancer remains a major cause of mortality worldwide.
One reason that bladder cancer is so serious is because, presently, detecting and treating bladder cancer is difficult.
However, because cystoscopy is an expensive and invasive procedure, it cannot be used as a general screening tool for the detection of bladder cancer.
Head and neck cancer remains a morbid and often fatal disease.
Detection of occult neoplastic cells in surrounding surgical margins is a strong predictor of local regional recurrence resulting in a significant decrease in overall survival
Where a DNA sequence is repetitive, genetic recombination can result in the loss of repeat DNA sequences or the gain of repeat DNA sequences at a genetic locus.
However, this diagnostic strategy has limited clinical application because the techniques are cumbersome and because p53 mutations appear relatively late in the disease.

Method used

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  • Method for detecting cell proliferative disorders
  • Method for detecting cell proliferative disorders
  • Method for detecting cell proliferative disorders

Examples

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example 1

Molecular Detection of Primary Bladder Cancer by Microsatellite DNA Analysis

[0073]The purpose of this Example is to show that microsatellite DNA markers are useful as clonal markers for the detection of human cancer, because simple DNA repeat mutations can be readily detected in clinical samples by the polymerase chain reaction. In this Example, the feasibility of polymerase chain reaction-based microsatellite DNA analysis of DNA from urine sediment is shown by correctly identifying nineteen of twenty patients with primary bladder tumors by this approach. In contrast, using uDne cytology, only nme of eighteen affected patients were detected.

[0074]Sixty trinucleotide and tetrucleotide markers in the DNA from fifty anonymous primary bladder cancers were screened. The screening was done in the following manner: Frozen tumor tissue was cut into 10 mu.m sections. All samples, including lymphocytes, were digested with 1% SDS-proteinase K at 60.degree. C. for 5 hr. DNA was extracted by eth...

example 2

Microsatellite DNA Alterations in Serum DNA of Head and Neck Cancer Patients

[0086]The purpose of this Example is to show that microsatellite DNA analysis of serum represents a novel method for the detection of circulating tumor cell DNA. Lymphocyte, serum and tumor DNA were retrospectively analyzed from twenty-one head and neck cancer patients. Head and neck cancer remains a morbid and often fatal disease. Large tumor bulk and tumor extension are predictors of local regional recurrence and poor outcome. Molecular detection of occult neoplastic cells in surrounding surgical margins is a strong predictor of local regional recurrence resulting in a significant decrease in overall survival. In this Example, twenty-one patients were followed from initial diagnosis of transitional cell carcinoma with microsatellite DNA analysis of urine DNA at the time of cystoscopic evaluations scheduled at routine intervals. In almost all cases, DNA-based analysis correlated precisely with clinical find...

example 3

Detection of Bladder Cancer Reoccurrence by Microsatellite DNA Analysis of Urine

[0097]The purpose of this Example was to demonstrate that the microsatellite DNA analysis method can be used for following-up patients with transitional cell carcinoma. A reliable, non-invasive method for monitoring patients with transitional cell carcinoma of the bladder would be of great clinical benefit. In this Example, serial urine samples were tested from twenty-one patients who had been treated for bladder cancer with twenty polymorphic microsatellite DNA markers in a blinded fashion. Recurrent lesions were detected in ten out of eleven patients and correctly predicted the existence of a neoplastic cell population in the urine of two patients, four and six months before cystoscopic evidence of the tumor. The assay was negative in ten of ten patients who had no evident cancer. This Example shows that microsatellite DNA analysis of urine sediment represents a novel and potentially powerful clinical ...

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Abstract

The present invention relates to the detection of a cell proliferative disorder associated with alterations of microsatellite DNA in a sample. The microsatellite DNA can be contained within any of a variety of samples, such as urine, sputum, bile, stool, cervical tissue, saliva, tears, or cerebral spinal fluid. The invention is a method to detect an allelic imbalance by assaying microsatellite DNA. Allelic imbalance is detected by observing an abnormality in an allele, such as an increase or decrease in microsatellite DNA which is at or corresponds to an allele. An increase can be detected as the appearance of a new allele. In practicing the invention, DNA amplification methods, particularly polymerase chain reactions, are useful for amplifying the DNA. DNA analysis methods can be used to detect such a decrease or increase. The invention is also a method to detect genetic instability of microsatellite DNA. Genetic instability is detected by observing an amplification or deletion of the small, tandem repeat DNA sequences in the microsatellite DNA which is at or corresponds to an allele. The invention is also a kit for practicing these methods.

Description

[0001]This application is a continuation of co-pending application Ser. No. 10 / 917,330 filed Aug. 13, 2004, which is a continuation application of 09 / 948,909 filed Sep. 10, 2001, now issued as U.S. Pat. No. 6,780,592, which is a continuation of Ser. No. 08 / 968,733 filed Aug. 28, 1997, now issued as U.S. Pat. No. 6,291,163, which claims the benefit of Provisional Application Ser. No. 60 / 025,805, filed Aug. 28, 1996.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The present invention relates generally to the detection of a target nucleic acid sequence and specifically to the detection of microsatellite DNA sequence mutations associated with a cell proliferative disorder.[0004]2. Description of Related Art[0005]Cancer remains a major cause of mortality worldwide. Despite advancements in diagnosis and treatment, the overall survival rate has not imrved significantly in the past twenty years. There remains an unfulfilled need for a more sensitive means of early diagnosis...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C12Q1/6883C12Q1/6886
CPCC12Q1/6883C12Q1/6886C12Q2600/118C12Q2600/112C12Q2600/158C12Q2600/172C12Q2600/156
Inventor SIDRANSKY, DAVID
Owner THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE
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